Erschienen in: Standards in Genomic Sciences ; 5 (2011), 3. - S. 298-310 Standards in Genomic Sciences (2011) 5:298-310 DOI:10.4056/sigs.2215005 Complete genome sequence of Parvibaculum T lavamentivorans type strain (DS-1 ) David Schleheck1*, Michael Weiss1, Sam Pitluck2, David Bruce3, Miriam L. Land4, Shunsheng Han3, Elizabeth Saunders3, Roxanne Tapia3, Chris Detter3, Thomas Brettin4, James Han2, Tanja Woyke2, Lynne Goodwin3, Len Pennacchio2, Matt Nolan2, Alasdair M. Cook1, Staffan Kjelleberg5, Torsten Thomas5 1 Department of Biological Sciences and Research School Chemical Biology, University of Konstanz, Germany 2 DOE Joint Genome Institute, Walnut Creek, California, USA 3 Los Alamos National Laboratory, Bioscience Division, Los Alamos, New Mexico, USA 4 Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA 5 Centre for Marine Bio-Innovation and School of Biotechnology and Biomolecular Science, University of New South Wales, Sydney, Australia *Corresponding author: [email protected] Keywords: Parvibaculum lavamentivorans DS-1, aerobic, Gram-negative, Rhodobiaceae, sur- factant biodegradation Parvibaculum lavamentivorans DS-1T is the type species of the novel genus Parvibaculum in the novel family Rhodobiaceae (formerly Phyllobacteriaceae) of the order Rhizobiales of Al- phaproteobacteria. Strain DS-1T is a non-pigmented, aerobic, heterotrophic bacterium and represents the first tier member of environmentally important bacterial communities that cata- lyze the complete degradation of synthetic laundry surfactants. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 3,914,745 bp long genome with its predicted 3,654 protein coding genes is the first com- pleted genome sequence of the genus Parvibaculum, and the first genome sequence of a rep- resentative of the family Rhodobiaceae. Introduction Parvibaculum lavamentivorans strain DS-1T P. lavamentivorans DS-1T is therefore an example (DSM13023 = NCIMB13966) was isolated for its of a first tier member of a two-step process that ability to degrade linear alkylbenzenesulfonate mineralizes environmentally important surfac- (LAS), a major laundry surfactant with a world- tants. wide use of 2.5 million tons per annum [1]. Strain Other representatives of the novel genus Parviba- DS-1T was difficult to isolate, is difficult to culti- culum have been recently isolated. Parvibaculum vate, and represents a novel genus in the Alpha- sp. strain JP-57 was isolated from seawater [6] proteobacteria [2,3]. Strain DS-1 catalyzes not only and is also difficult to cultivate [3]. Parvibaculum the degradation of LAS, but also of 16 other com- indicum sp. nov. was also isolated from seawater, mercially important anionic and non-ionic surfac- via an enrichment culture that degraded polycyc- tants (hence the species name lavamentivorans = lic aromatic hydrocarbons (PAH) and crude oil [7]. consuming [chemicals] used for washing [3]). The Another Parvibaculum sp. strain was isolated from initial degradation as catalyzed by strain DS-1T a PAH-degrading enrichment culture, using river involves the activation and shortening of the alkyl- sediment as inoculum [8]. Parvibaculum species chain of the surfactant molecules, and the excre- were also reported in a study on marine alkane- tion of short-chain degradation intermediates. degrading bacteria [9]. Parvibaculum species are These intermediates are then completely utilized frequently detected by cultivation-independent by other bacteria in the community [4,5]. methods, predominantly in habitats or settings The Genomic Standards Consortium Konstanzer Online-Publikations-System (KOPS) URL: http://nbn-resolving.de/urn:nbn:de:bsz:352-177182 Schleheck et al. with hydrocarbon degradation. These include a succinate, or alkanes, alkanols and alkanoates (C8 - bacterial community on marine rocks polluted C16); no sugars tested were utilized [3]. with diesel oil [10], a bacterial community from To allow for growth in liquid culture with most of diesel-contaminated soil [11], a petroleum- the 16 different surfactants at high concentrations degrading bacterial community from seawater (e.g. for LAS, >1 mM; see [3].), the culture fluid [12], an oil-degrading cyanobacterial community needs to be supplemented with a solid surface, e.g. [13] and biofilm communities in pipes of a district polyester fleece or glass fibers [2,3]. The addition- heating system [14]. Parvibaculum species have al solid surface is believed to support biofilm for- also been detected in denitrifying, linear- mation, especially in the early growth phase when nonylphenol (NP) degrading enrichment cultures the surfactant concentration is high, and the or- from NP-polluted river sediment [15] and in ganism grows as single, suspended cells (non- groundwater that had been contaminated by li- motile) during the later growth phase. Growth near alkyl benzenes (LABs; non-sulfonated LAS] with a non-membrane toxic substrate (e.g. ace- [16]. Additionally, Parvibaculum species were tate) is independent of a solid surface, and consti- detected in biofilms that degraded polychlori- tutes suspended, single cells (motile). We presume nated biphenyls (PCBs) using pristine soil as in- that the biofilm formation by strain DS-1T is a pro- oculum [17], and in a PAH-degrading bacterial tective response to the exposure to membrane- community from deep-sea sediment of the West solubilizing agents (cf. [30]). Pacific [18]. Finally, Parvibaculum species were Based on the 16S rRNA gene sequence, strain DS1T detected in an autotrophic Fe(II)-oxidizing, ni- was described as the novel genus Parvibaculum, trate-reducing enrichment culture [19], as well as which was originally placed in the family Phyllo- in Tunisian geothermal springs [20]. The wide- bacteriaceae within the order Rhizobiales of Al- spread occurrence of Parvibaculum species in ha- phaproteobacteria [3,31]. The nearest well- bitats or settings related to hydrocarbon degrada- described organism to strain DS-1T is Afifella ma- tion implies an important function and role of rina (formerly Rhodobium marinum) (92% 16S these organisms in environmental biodegradation, rRNA gene sequence identity), a photosynthetic despite their attribute as being difficult to culti- purple, non-sulfur bacterium. The genus Rhodo- vate in a laboratory. bium was later re-classified as a member of the Here we present a summary classification and a novel family Rhodobiaceae [26,32], together with set of features for P. lavamentivorans DS-1T, to- two novel genera of other photosynthetic purple gether with the description of a complete genome non-sulfur bacteria (Afifella and Roseospirillum), sequence and annotation. The genome sequencing as well as with two novel genera of heterotrophic and analysis was part of the Microbial Genome aerobic bacteria, represented by the red- Program of the DOE Joint Genome Institute. pigmented Anderseniella baltica (gen. nov., sp. nov.) [33,34] and non-pigmented Tepidamorphus Classification and features gemmatus (gen. nov., sp. nov.) [35,36]. A phyloge- P. lavamentivorans DS-1T is a Gram-negative, non- netic tree (Figure 2) was constructed with the 16S pigmented, very small (approx. 1.0 × 0.2 µm), rRNA gene sequence of P. lavamentivorans DS-1T slightly curved rod-shaped bacterium that can be and that of (i) other isolated Parvibaculum strains, motile by means of a polar flagellum (Figure 1, (ii) representatives of other genera within the Table 1). Strain DS-1T grows very slowly on com- family Rhodobiaceae, (iii) representatives of the plex medium (e.g. on LB- or peptone-agar plates) genera in the family Phyllobacteriaceae, as well as, and forms pinpoint colonies only after more than (iv) representatives of other families within the two weeks of incubation. The organism can be order Rhizobiales. The phylogenetic tree shows quickly overgrown by other organisms. Larger now the placement of Parvibaculum species within colonies are obtained when the complex medium the family Rhodobiaceae, and that the Parvibacu- is supplemented with a surfactant, e.g. Tween 20 lum sequences clustered as a distinct evolutionary (see DSM-medium 884 [29]) or LAS [3]. When lineage within this family (Figure 2). This classifi- cultivated in liquid culture with mineral-salts me- cation of Parvibaculum has been adopted in the dium, strain DS-1T grows within one week with Ribosomal Database Project (RDP) and SILVA the single carbon sources acetate, ethanol, or rRNA Database Project, but not in the GreenGenes database. The family Rhodobiaceae has also not http://standardsingenomics.org 299 Parvibaculum lavamentivorans type strain (DS-1T) been included in the NCBI-taxonomy, IMG- phosphatidyl glycerol, diphosphatidyl glycerol, taxonomy, and GOLD databases. phosphatidyl ethanolamine, phosphatidyl choline, Currently, 360 genome sequences of members of and two, unidentified aminolipids; the presence of the order Rhizobiales of Alphaproteobacteria have the two additional aminolipids appears to be dis- been made available (GOLD database; August tinctive of the organism [3]. The G+C content of 2011), and within the family Phyllobacteriaceae the DNA was determined to be 64% [3], which there are 21 genome sequences available corresponds well to the G+C content observed for (Chelativorans sp. BNC1, Hoeflea phototrophica the complete genome sequence (see below). DFL-43, and 18 Mesorhizobium strains). No ge- nome sequences currently exist for a representa- Genome sequencing information tive of the novel family Rhodobiaceae, except